The Dynamic Changes of Brassica napus Seed Microbiota across the Entire Seed Life in the Field.

The seed microbiota is an important component given by nature to plants, protecting seeds from damage by other organisms and abiotic stress. However, little is known about the dynamic changes and potential functions of the seed microbiota during seed development. In this study, we investigated the composition and potential functions of the seed microbiota of rapeseed (Brassica napus). A total of 2496 amplicon sequence variants (ASVs) belonging to 504 genera in 25 phyla were identified, and the seed microbiota of all sampling stages were divided into three groups. The microbiota of flower buds, young pods, and seeds at 20 days after flowering (daf) formed the first group; that of seeds at 30 daf, 40 daf and 50 daf formed the second group; that of mature seeds and parental seeds were clustered into the third group. The functions of seed microbiota were identified by using PICRUSt2, and it was found that the substance metabolism of seed microbiota was correlated with those of the seeds. Finally, sixty-one core ASVs, including several potential human pathogens, were identified, and a member of the seed core microbiota, Sphingomonas endophytica, was isolated from seeds and found to promote seedling growth and enhance resistance against Sclerotinia sclerotiorum, a major pathogen in rapeseed. Our findings provide a novel perspective for understanding the composition and functions of microbiota during seed development and may enhance the efficiency of mining beneficial seed microbes.

Protective effect of Tibetan medicine Qiwei Tiexie pills on liver injury induced by acetaminophen overdose: An integrated strategy of network pharmacology, metabolomics and transcriptomics.

BACKGROUND: Drug-induced liver injury, particularly from acetaminophen (APAP), has emerged as a significant public health concern. Unfortunately, there is currently no effective treatment strategy available. Qiwei Tiexie pills (QWTX), a traditional Tibetan medicine, have demonstrated considerable clinical efficacy in treating various liver diseases. Nevertheless, the protective effect of QWTX against drug-induced liver injury and its underlying mechanism remains poorly understood. PURPOSE: This study aimed to assess the therapeutic potential of QWTX, a Tibetan medicine, in an animal model of APAP-induced liver injury. Additionally, we sought to investigate the molecular mechanism through which QWTX exerts its effects. METHODS: We employed LC-MS and network pharmacology to predict the potential targets of QWTX in drug-induced liver injury. Subsequently, we employed HE staining, transcriptomics, metabolomics, and qRT-PCR to analyze the mechanism underlying QWTX treatment in drug-induced liver injury. RESULTS: Network pharmacology analysis revealed that the active components of QWTX are involved in inflammatory and drug metabolism-related pathways. In mouse models, pretreatment with QWTX effectively mitigated the elevated levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and inflammatory factors (IL-1ß, IL-6, and TNF-α) induced by APAP overdose. Moreover, APAP inhibited 1459 differentially expressed genes (DEGs) and 874 differential accumulation metabolites (DAMs), while QWTX promoted their expression. Conversely, APAP promoted 874 genes and 119 metabolites, which were inhibited by QWTX. Further analysis demonstrated that QWTX ameliorated the metabolic disorders induced by APAP overdose and potentially exerted a protective effect by inhibiting the expression of critical genes in crucial inflammatory pathways. QWTX also up-regulated antioxidant enzymes, thereby mitigating the oxidative stress resulting from APAP overdose. CONCLUSION: QWTX treatment effectively protects against APAP-induced liver damage in mice. Transcriptomic and metabolomic analyses further revealed that QWTX ameliorated hepatic metabolic disorders induced by APAP overdose while significantly suppressing the inflammatory response and oxidative stress associated with drug-induced liver injury. This study provides a new insight into the treatment of drug-induced liver injury by the TCM system and provides a basis for the development of new therapies for drug-induced liver injury by QWTX and its active ingredients.

Editing homologous copies of an essential gene affords crop resistance against two cosmopolitan necrotrophic pathogens.

Sclerotinia sclerotiorum and Botrytis cinerea are typical necrotrophic pathogens that can attack more than 700 and 3000 plant species, respectively, and cause huge economic losses across numerous crops. In particular, the absence of resistant cultivars makes the stem rot because of S. sclerotiorum the major threat of rapeseed (Brassica napus) worldwide along with Botrytis. Previously, we identified an effector-like protein (SsSSVP1) from S. sclerotiorum and a homologue of SsSSVP1 on B. cinerea genome and found that SsSSVP1 could interact with BnQCR8 of rapeseed, a subunit of the cytochrome b-c1 complex. In this study, we found that BnQCR8 has eight homologous copies in rapeseed cultivar Westar and reduced the copy number of BnQCR8 using CRISPR/Cas9 to improve rapeseed resistance against S. sclerotiorum. Mutants with one or more copies of BnQCR8 edited showed strong resistance against S. sclerotiorum and B. cinerea. BnQCR8-edited mutants did not show significant difference from Westar in terms of respiration and agronomic traits tested, including the plant shape, flowering time, silique size, seed number, thousand seed weight and seed oil content. These traits make it possible to use these mutants directly for commercial production. Our study highlights a common gene for breeding of rapeseed to unravel the key hindrance of rapeseed production caused by S. sclerotiorum and B. cinerea. In contrast to previously established methodologies, our findings provide a novel strategy to develop crops with high resistance against multiple pathogens by editing only a single gene that encodes the common target of pathogen effectors.

Effects of medicine food Fructus Gardeniae on liver and kidney functions after oral administration to rats for 12 weeks.

Fructus Gardeniae (FG) is medicine food widely used for the treatment and prevention of various diseases. However, in recent years, research has suggested that high doses of FG can cause hepatotoxicity and nephrotoxicity. To assess this potential toxicity in more depth, this study investigated the effects of decocted FG and two of its bioactive constituents (geniposide and genipin) on liver and kidney function in rats. Rats were intragastrically administered FG (330 mg/kg body weight), geniposide (50 mg/kg body weight), or genipin (50 mg/kg body weight) for 12 weeks. Changes in body weight, liver and kidney indices, biochemical indices, and inflammatory factors were monitored. In addition, pathological sections were assessed and the expression of caspase-3, NF-κBp65, COX-2, and iNOS was detected by immunohistochemistry and Western blot. It was found that the levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, creatinine, and urea nitrogen increased following administration of FG, geniposide, and genipin. Furthermore, the activities of superoxide dismutase and reduced glutathione decreased following treatment, while malondialdehyde levels increased. Pathological and immunohistochemical evaluations further confirmed that FG and its constituents may cause damage to the liver and kidneys. The mechanism study revealed that the protein level of inflammatory pathway increased and further promoted apoptosis, suggesting that it should not be taken orally for extended periods of time. PRACTICAL APPLICATIONS: Chinese medicine and food safety have always been public health concerns. Fructus Gardeniae (FG) is a plant with a dual-purpose as it is used as both a medicine and food. Medicinally, it has the effects of heat-clearing and detoxification. However, its adverse effects and related mechanisms are not clear, and this has potential safety implications. In this study, rats were treated with FG for 12 weeks and found that the long-term administration of FG or high dosing can lead to damage to liver and kidney function. Therefore, close attention must be paid to the dosage of FG in order to achieve a therapeutic effect and avoid adverse reactions. Thus, this study lays a foundation for the safety evaluation and clinical application of FG.

Four Novel Botourmiaviruses Co-Infecting an Isolate of the Rice Blast Fungus Magnaporthe oryzae.

Via virome sequencing, six viruses were detected from Magnaporthe oryzae strains YC81-2, including one virus in the family Tombusviridae, one virus in the family Narnaviridae and four viruses in the family Botourmiaviridae. Since the RNA-dependent RNA polymerase (RdRp) of one botourmiavirus show the highest identity (79%) with Magnaporthe oryzae ourmia-like virus 1 (MOLV1), the virus that was grouped into the genus Magoulivirus was designated as Magnaporthe oryzae botourmiavirus 2 (MOBV2). The three other novel botourmiaviruses were selected for further study. The complete nucleotide sequences of the three botourmiaviruses were determined. Sequence analysis showed that virus 1, virus 2, and virus 3 were 2598, 2385, and 2326 nts in length, respectively. The variable 3' untranslated region (3'-UTR) and 5'-UTR of each virus could be folded into a stable stem-loop secondary structure. Each virus consisted of a unique ORF encoding a putative RdRp. The putative proteins with a conserved GDD motif of RdRp showed the highest sequence similarity to RdRps of viruses in the family Botourmiaviridae. Phylogenetic analysis demonstrated that these viruses were three distinct novel botourmiaviruses, clustered into the Botourmiaviridae family but not belonging to any known genera of this family. Thus, virus 1, virus 2, and virus 3 were designated as Magnaporthe oryzae botourmiavirus 5, 6, and 7 (MOBV5, MOBV6, and MOBV7), respectively. Our results suggest that four distinct botourmiaviruses, MOBV2, MOBV5, MOBV6, and MOBV7, co-infect a single strain of Magnaporthe oryzae, and MOBV5, MOBV6, and MOBV7 are members of three unclassified genera in the family Botourmiaviridae.

Defective modulation of LINE-1 retrotransposition by cancer-associated SAMHD1 mutants.

Long interspersed nuclear elements (LINE-1) is now considered as the only active autonomous mobile DNA in humans, LINE-1 retrotransposition activities are associated with and fluctuate during cancer initiation and progression; however, the mechanism underlying the increased LINE-1 activity in cancer is poorly understood. SAMHD1 has been reported to be a potent inhibitor of LINE-1 retrotransposition, and SAMHD1 mutations are frequently associated with cancer development. To gain insights on whether cancer-related SAMHD1 mutants affect LINE-1 activity, we explored the biochemical and cellular properties of some human mutants known correlate with the development of cancer. Most of the tested SAMHD1 cancer-related mutations were defective in LINE-1 inhibition. Interestingly we also found that SAMHD1 mutant K288T was defective for dNTPase activity but showed potent activity against LINE-1 retrotransposition. These findings suggest that LINE-1 inhibition does not depend solely on the dNTPase activity of SAMHD1. In contrast, SAMHD1's ability to inhibit ORF2p-mediated LINE-1 RNP reverse transcription was correlated with SAMHD1-mediated LINE-1 inhibition. Together, our data could also facilitate the deeper understanding for the inhibition of endogenous LINE-1 elements by SAMHD1.

The research advances of exosomes in esophageal cancer.

Esophageal carcinoma (EC) is one of the most common human digestive tract tumors, with high morbidity and mortality. It is necessary to elucidate the mechanism of cancer progression and seek early EC diagnostic markers for prompt detection and intervention. Exosomes are membrane nanovesicles secreted from many nucleated cells, 30-100 nm in diameter, containing various proteins, lipids and nucleic acids. They exist in peripheral blood, urine, ascites and other body fluids, widely engaged with intercellular material exchange and signal communication. Exosomes secreted from EC cells or tissues conduct important functions in tumor growth and progression. The detection and analysis of tumor-derived or tumor-associated exosomes has potential for EC early diagnosis and prognosis assessment. In the present paper, the exosomes' biological behaviors, isolation, detection and functions in EC progression - using as potential biomarkers for EC diagnosis or prognosis - are reviewed.

A scientometric analysis on hepatocellular carcinoma magnetic resonance imaging research from 2008 to 2017.

BACKGROUND: With the development of new magnetic resonance imaging (MRI) techniques, an increasing number of articles have been published regarding hepatocellular carcinoma magnetic resonance imaging (HCCMRI) in the past decade. However, few studies have statistically analyzed these published articles. In this study, we aim to systematically evaluate the scientific outcomes of HCCMRI research and explore the research hotspots from 2008 to 2017. METHODS: The included articles regarding HCCMRI research from 2008 to 2017 were downloaded from the Web of Science Core Collection and verified by two experienced radiologists. Excel 2016 was used to analyze the literature data, including the publication years and journals. CiteSpace V was used to perform co-occurrence analyses for authors, countries/regions and institutions and to generate the related collaboration network maps. Reference co-citation analysis (RCA) and burst keyword detection were also performed using CiteSpace V to explore the research hotspots in the past decade. RESULTS: A total of 835 HCCMRI articles published from 2008 to 2017 were identified. Journal of Magnetic Resonance Imaging published the most articles (79 publications, 9.46%). Extensive cooperating relationship were observed among countries/regions and among authors. South Korea had the most publications (199 publications, 21.82%), followed by the United States of America (USA) (190 publications, 20.83%), Japan (162 publications, 17.76%), and the People's Republic of China (148 publications, 16.23%). Among the top 10 co-cited authors, Bruix J (398 citations) was ranked first, followed by Llovet JM (235 citations), Kim YK (170 citations) and Forner A (152 citations). According to the RCA, ten major clusters were explored over the last decade; "LI-RADS data system" and "microvascular invasion" (MVI) were the two most recent clusters. Forty-seven burst keywords with the highest citation strength were detected over time. Of these keywords, "microvascular invasion" had the highest strength in the last 3 years. The LI-RADS has been constantly updated with the latest edition released in July 2018. However, the LI-RADS still has limitations in identifying certain categories of lesions by conceptual and non-quantitative probabilistic methods. Plenty of questions still need to be further answered such as the difference of diagnostic efficiency of each major/ancillary imaging features. Preoperative prediction of MVI of HCC is very important to therapeutic decision-making. Some parameters of Gd-EOB-DTPA-enhanced MRI were found to be useful in prediction of MVI, however, with a high specificity but a very low sensitivity. Comprehensive predictive model incorporating both imaging and clinical variables may be the more preferable in prediction of MVI of HCC. CONCLUSIONS: HCCMRI-related publications displayed a gradually increasing trend from 2008 to 2017. The USA has a central position in collaboration with other countries/regions, while South Korea contributed the most in the number of publications. Of the ten major clusters identified in the RCA, the two most recent clusters were "LI-RADS data system" and "microvascular invasion", indicative of the current HCCMRI research hotspots.

The Possible Mechanisms of HSV-TK/Hyperthermia Combined with 131I-antiAFPMcAb-GCV Nanospheres to Treat Hepatoma.

Our previous findings showed a good therapeutic effect of the combination of suicide gene HSV-TK, nuclide 131I, and magnetic fluid hyperthermia (MFH) on hepatoma by using magnetic nanoparticles as linkers, far better than any monotherapy involved, with no adverse effects. This combination therapy might be an eligible strategy to treat hepatic cancer. However, it is not clear how the combination regimen took the therapeutic effects. In the current study, to explore the possible mechanisms of radionuclide-gene therapy combined with MFH to treat hepatoma at tissue, cellular, and molecular levels and to provide theoretical and experimental data for its clinical application, we examined the apoptosis induction of the combination therapy and investigated the expression of the proteins related to apoptosis such as survivin, livin, bcl-2, p53, and nucleus protein Ki67 involved in cell proliferation, detected VEGF, and MVD involved in angiogenesis of tumor tissues and analyzed the pathologic changes after treatment. The results showed that the combination therapy significantly induced the hepatoma cell apoptosis. The expression of survivin, VEGF, bcl-2, p53, livin, Ki67, and VEGF proteins and microvascular density (MVD) were all decreased after treatment. The therapeutic mechanisms may be involved in the downregulation of Ki67 expression leading to tumor cell proliferation repression and inhibition of survivin, bcl-2, p53, and livin protein expression inducing tumor cell apoptosis, negatively regulating VEGF protein expression, and reducing vascular endothelial cells, which results in tumor angiogenesis inhibition and microvascular density decrease and tumor cell necrosis. These findings offer another basic data support and theoretical foundation for the clinical application of the combination therapy.

Cumulative dose of radiation therapy of hepatocellular carcinoma patients and its deterministic relation to radiation-induced liver disease.

This study aimed to investigate the relationship between dose and radiation-induced liver disease (RILD) in patients with hepatocellular carcinoma (HCC) receiving 3-dimensional conformal radiotherapy (3DCRT). Twenty-three patients with HCC who received conventional fractionated 3DCRT, including 7 who were diagnosed with classic RILD, were enrolled in this retrospective investigation. Cone-beam computed tomography (CBCT) scans were acquired at the time of treatment for each patient. The beams from each patient's treatment plan were applied to each pretreatment CBCT (the modified CBCT or mCBCT) to construct the delivered dose distribution of the day considering inter-treatment anatomy changes. The daily doses were summed together with the help of deformable image registration (DIR) to obtain the adjusted cumulative dose (Dadjusted). The dose changes to the normal liver between the original planned dose (Dplan) and Dadjusted were evaluated by V20, V30, V40, and the mean dose to normal liver (MDTNL). Univariate analysis was performed to identify the significant dose changes. Among the 23 patients, the liver V20, V30, V40, and MDTNL showed significant differences between Dplan and Dadjusted, with average values of these parameters increased by 4.1%, 4.7%, 4.5%, and 3.9 Gy, respectively (p < 0.05). The adjusted liver dose in 21 patients (91%) was higher than the planned value. For patients without and with RILD,the MDTNL was increased on average by 3.5 Gy and 4.7 Gy, and normal tissue complication probability (NTCP) increased on average by 2.8% and 7.5%, respectively. Our study found that the adjusted cumulative dose based on calculations using pretreatment mCBCT differs significantly from planned dose; the use of the dosimetric results of the initial plan was found to be less predictive of RILD as compared with Dadjusted. Determination of a reconstructed Dadjusted using the mCBCT scans are more accurate in predicting RILD and has the potential to reduce the risk of RILD.

The Recent Advances on Liver Cancer Stem Cells: Biomarkers, Separation, and Therapy.

As the third major reason of mortality related to cancer in the world, liver cancer is also the fifth most frequent cancer. Unluckily, a majority of patients succumb and relapse though many progresses have been made in detection and therapy of liver cancer. It has been put forward that in liver cancer, cancer stem cells (CSCs) hold main responsibility for the formation, invasion, metastasis, and recurrence of tumor. Strategies that are intended to target liver CSCs are playing a more and more significant role in supervising the development of liver cancer treatment and assessing new therapeutic methods. Herein, a brief review about molecule markers, signal pathways, separation, and treatment on liver cancer stem cells (LCSCs) is provided in this paper.

Antibiotic use in rural China: a cross-sectional survey of knowledge, attitudes and self-reported practices among caregivers in Shandong province.

BACKGROUND: To improve antibiotic use globally, we must deepen our understanding of the public's knowledge, attitudes and practices (KAP) concerning antibiotics. Children are frequent users of antibiotics, and their caregivers play important roles in determining how antibiotics are used. The purpose of this study was to describe caregivers' KAP in a rural province in eastern China, and to identify socio-demographic factors associated with inappropriate antibiotic use. METHODS: A cross-sectional questionnaire based survey was conducted in 12 villages in one county in Shandong Province. A total of 727 individuals who were the primary day-to-day caregiver for a child aged 0-7 years were randomly selected and invited to participate. All caregivers were surveyed face-to-face using a semi-structured questionnaire focusing on the use of antibiotics in children. RESULTS: Almost all invited caregivers (99.3 %) completed the questionnaire in full. Caregivers expressed high levels of over-expectation for antibiotics for common childhood symptoms, stating that antibiotics were always or usually necessary when a child has a fever (46 %) or dry cough (42 %). Most caregivers (93 %) were aware that they should follow the doctor's advice when giving their children antibiotics. Many, however, reported that they had previously deviated from advice; this was most commonly through using antibiotics intermittently rather than regularly, but also by increasing and decreasing doses. Caregivers that were older and that had less formal education had higher levels of self-reported adherence (p < 0.01). A third of caregivers admitted to storing leftover antibiotics at home, and almost all of these caregivers (97 %) had used the antibiotics on a second occasion for their child. CONCLUSION: We have identified important gaps in knowledge, attitudes and practices concerning antibiotics among this rural population of caregivers. There is a clear need for multifaceted interventions that target village doctors, to improve prescribing and communication, as well as the general public, to improve health-seeking behaviours and promote responsible individual use of antibiotics.

[Three-dimensional reconstruction and digitized visualization of the sacrum of women of Uygur and Han nationalities in Xinjiang].

OBJECTIVE: To construct digitized three-dimensional models of the sacrum of women of Uygur and Han nationalities in Xinjiang and assess their value in anatomical studies. METHODS: Ninety Uygur women and 90 Han women of child-bearing age in Xinjiang were recruited. Digitized three-dimensional reconstruction was used to construct the sacrum models for morphological study and measurement of the physiological indices. RESULTS: The sacral forms of Han women were consistent with the standard anatomical descriptions. Of the Uygur women, 57 (63.3%) showed a standard type of the sacrum, 6 (6.7%) had 4 pairs of sacral foramens, 9 (10%) had 5 pairs of sacral foramens, and 18 (20%) had sacrococcygeal joint abnormalities. There were significant differences in sacral morphologies between the two nationalities (P<0.05). The depths of the sacrum foramen and sacral hiatus were significantly greater in Uyghur women than in Han women (P<0.05). Only a minor proportion of the cases had nonstandard sacral types, found only in Uygur women. CONCLUSIONS: Uygur and Han women of child-bearing age in Xinjiang have morphological and anatomical differences of the sacrum, which should be considered in clinical practice.

Tributyltin chloride-induced immunotoxicity and thymocyte apoptosis are related to abnormal Fas expression.

Tributyltin chloride (TBTC), a characteristic organotin compound, is widely used as an agricultural pesticide, as a stabiliser for polyvinyl-chloride plastics and in antifouling paints for ship hulls. Organotin compounds are known to produce toxicity in the immune system, but the mechanism underlying this immunotoxicity remains unclear. In this study, we evaluated the immunotoxic effect of TBTC on the acquired immune response, and we investigated the involvement of thymocyte apoptosis and Fas expression in the observed immunotoxicity of TBTC. Mice were randomly assigned to four groups (10 mice per group) and treated with TBTC at doses of 0, 0.5, 4 and 20 mg/kg by oral gavage for 28 days. Following TBTC administration, animals were sacrificed, and morphologic changes in the thymus and spleen were assessed. Atrophy in both the thymus and spleen was observed in all groups treated with TBTC, and the relative organ weight in the highest TBTC group (20 mg/kg) was significantly lower than that observed in the control group. We also conducted assays to assess the cellular and humoral functional responses such as plaque-forming cell assay (PFC), lymphocyte proliferation test and delayed-type hypersensitivity (DTH) response to SRBC. Our results indicate that at doses of 4 mg/kg and 20 mg/kg, TBTC could significantly suppress both the humoral and cellular immune responses when compared to the control group (p<0.05). In addition, immunohistochemical staining and flow cytometry analysis were carried out to measure the expression of Fas and thymocyte apoptosis, respectively. We observed a dose-dependent increase in thymocyte apoptosis and that Fas expression in the TBTC-treated groups (4 mg/kg and 20 mg/kg) was significantly enhanced when compared to the control group. Correlation analysis demonstrated a positive linear correlation between apoptosis and Fas expression, indicating that TBTC-induced thymocyte apoptosis may be mediated by Fas expression. Taken together, our data clearly demonstrate that TBTC-induced immunotoxicity is associated with thymocyte apoptosis and that this process is mediated by the Fas pathway.

[Analysis on incidence of bacillary dysentery in Jinan municipality from 1951 to 2005].

OBJECTIVE: To explore the epidemic characteristics of bacterial dysentery in Jinan municipality, and to provide scientific basis for effective strategy for bacterial dysentery control. METHODS: The epidemiological characteristics of bacillary dysentery in Jinan from 1951 to 2005 were analyzed. A total of 485,333 cases in the span of 50 years were recorded, while the population-based case distribution was less than the total cases due to the data incompleteness during the Cultural Revolution. RESULTS: The incidence of bacillary dysentery in Jinan has been decreasing by years with average incidence rate of 283.10/100,000. The significant differences were observed among the incidence rates of various ages(chi2 = 14.99, P < 0.05). There were four epidemic peaks, and all the incidence rates were about 1000/100,000. Age of onset mainly concentrated in the 0-4 years old, 20-years old and 30-years old. In terms of occupational distribution, workers accounted for 30.31%, the living-scattered children accounted for 22.71%, and the farmers accounted for 17.90%. The incidence focus was from July to September, which accounted for 71.57%. The peak of incidence emerged in August. The highest incidence in urban was 550.94/100,000. CONCLUSION: Through the efforts of several generations of health workers, the incidence of bacillary dysentery in Jinan has been basically brought under control. Further step should be taken for the control of bacterial dysentery in urban areas and the management of bacterial dysentery in rural areas. Moreover, the biological characteristics of F2a should be a focus for the future study.